title: Altered Ca2+ concentration, permeability and buffering in the myofibre Ca2+ store of a mouse model of malignant hyperthermia.
abstract: Malignant hyperthermia (MH) is linked to mutations in the type 1 ryanodine receptor, RyR1, the Ca2+ channel of the sarcoplasmic reticulum (SR) of skeletal muscle. The Y522S MH mutation was studied for its complex presentation, which includes structurally and functionally altered cell 'cores'. Imaging cytosolic and intra-SR [Ca2+] in muscle cells of heterozygous YS mice we determined Ca2+ release flux activated by clamp depolarization, permeability (P) of the SR membrane (ratio of flux and [Ca2+] gradient) and SR Ca2+ buffering power (B). In YS cells resting [Ca2+]SR was 45% of the value in normal littermates (WT). P was more than doubled, so that initial flux was normal. Measuring [Ca2+]SR(t) revealed dynamic changes in B(t). The alterations were similar to those caused by cytosolic BAPTA, which promotes release by hampering Ca2+-dependent inactivation (CDI). The [Ca2+] transients showed abnormal 'breaks', decaying phases after an initial rise, traced to a collapse in flux and P. Similar breaks occurred in WT myofibres with calsequestrin reduced by siRNA; calsequestrin content, however, was normal in YS muscle. Thus, the Y522S mutation causes greater openness of the RyR1, lowers resting [Ca2+]SR and alters SR Ca2+ buffering in a way that copies the functional instability observed upon reduction of calsequestrin content. The similarities with the effects of BAPTA suggest that the mutation, occurring near the cytosolic vestibule of the channel, reduces CDI as one of its primary effects. The unstable SR buffering, mimicked by silencing of calsequestrin, may help precipitate the loss of Ca2+ control that defines a fulminant MH event.
authors.affiliation: S. L. Hamilton: susanh@bsm.edu; E. Ríos: Rush University School of Medicine, Department of Molecular Biophysics and Physiology, 1750 West Harrison St., Suite 1279JS, Chicago, IL 60612, USA. Email: erios@rush.edu.
authors.forename: Carlo
authors.initials: C
authors.lastname: Manno
authors.forename: Lourdes
authors.initials: L
authors.lastname: Figueroa
authors.forename: Leandro
authors.initials: L
authors.lastname: Royer
authors.forename: Sandrine
authors.initials: S
authors.lastname: Pouvreau
authors.forename: Chang Seok
authors.initials: CS
authors.lastname: Lee
authors.forename: Pompeo
authors.initials: P
authors.lastname: Volpe
authors.forename: Alessandra
authors.initials: A
authors.lastname: Nori
authors.forename: Jingsong
authors.initials: J
authors.lastname: Zhou
authors.forename: Gerhard
authors.initials: G
authors.lastname: Meissner
authors.forename: Susan L
authors.initials: SL
authors.lastname: Hamilton
authors.forename: Eduardo
authors.initials: E
authors.lastname: Ríos
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism; D002155:Calsequestrin; D004195:Disease Models, Animal; D007035:Hypothermia / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009154:Mutation; D012519:Sarcoplasmic Reticulum / Q000378:metabolism*
chemical_list: D002135:Calcium-Binding Proteins; D002155:Calsequestrin; C503130:Casq1 protein, mouse; D002118:Calcium